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Gene and application, gene element, method for synthesizing odd-numbered medium chain fatty aldehyde and method for synthesizing even-numbered medium chain fatty hydrocarbon

A technology of gene element and fatty aldehyde, applied in the field of synthesizing even-numbered medium-chain aliphatic hydrocarbons, to achieve the effect of reducing the burden

Inactive Publication Date: 2016-05-18
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But there is no work to directly regulate the downstream synthetic pathway

Method used

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  • Gene and application, gene element, method for synthesizing odd-numbered medium chain fatty aldehyde and method for synthesizing even-numbered medium chain fatty hydrocarbon
  • Gene and application, gene element, method for synthesizing odd-numbered medium chain fatty aldehyde and method for synthesizing even-numbered medium chain fatty hydrocarbon
  • Gene and application, gene element, method for synthesizing odd-numbered medium chain fatty aldehyde and method for synthesizing even-numbered medium chain fatty hydrocarbon

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1: Construction of Genetic Elements for the Synthesis of Odd Medium Chain Fatty Aldehydes

[0069] Experimental Materials:

[0070] Thioesterase (TesA'): Escherichia coli endogenous thioesterase gene I (tesA') was purchased from addgene (Plasmid24636), and the gene was placed on a plasmid with a replicon of p15A and a promoter of placUV5, named pKS1, And the 75bp nucleotide after the initiation codon ATG was removed. The amino acid sequence encoded by the removed nucleotides is a signal peptide for localizing the enzyme in the intercellular space. Removing this signal peptide sequence can enrich the thioesterase in the cell and make E. coli produce a large amount of free fatty acid.

[0071] α-dioxygenase (Dox): According to the reported protein sequence of α-dioxygenase (NCBI Reference Sequence: NP_001066718.1) in rice (Oryzasativa), codon optimization was carried out according to Escherichia coli, and the optimized coding α- The DNA molecule of dioxygenase ...

Embodiment 2

[0091] Example 2: Construction of gene elements for the synthesis of odd-numbered medium-chain fatty alcohols

[0092] Experimental Materials:

[0093] AdhE: Aldehyde-CoA reductase / iron ion-dependent alcohol dehydrogenase, from E.coliBL21(DE3) genome (NCBI-GeneID: 8180074), a single point mutation replaced the NcoI restriction site in the sequence.

[0094] AdhP: Alcohol active dehydrogenase / acetaldehyde active reductase, from E. coliBL21(DE3) genome (NCBI-GeneID: 8181169).

[0095] YqdD: NADPH-dependent acetaldehyde reductase, from the E.coliBL21 (DE3) genome (NCBI-GeneID: 8180496), a single point mutation replaced the NdeI restriction site in the sequence.

[0096] YjgB: Alcohol dehydrogenase (atypical zinc-type alcohol dehydrogenase-like protein, zinc and NADPH dependent), from E. coliBL21(DE3) genome (NCBI-GeneID: 8182107).

[0097] Slr1192: Alcohol dehydrogenase containing zinc, derived from Synechocystis sp. PCC6803 strain (Synechocystissp.PCC6803), the protein sequenc...

Embodiment 3

[0125] Example 3: Construction of gene elements for the synthesis of even-numbered medium-chain aliphatic hydrocarbons

[0126] Experimental Materials:

[0127] CER1: Fatty aldehyde decarbonylase, derived from Arabidopsis thaliana (Arabidopsisthaliana), the protein sequence is UniProtKB / Swiss-Prot:F4HVY0.1, codon optimization is carried out according to Escherichia coli, and the optimized DNA molecule encoding CER1 has SEQ ID NO:3 Nucleotide sequences shown, genes synthesized in genscript.

[0128] AD9313: Fatty aldehyde decarbonylase, derived from Prochlorococcus marinus (Prochlorococcus marinus MIT9313), the protein sequence is NCBIReferenceSequence: NP_895059.1, codon optimization is carried out according to Escherichia coli, and the DNA molecule encoding AD9313 after optimization has SEQ ID NO: 4 Nucleotide sequences, genes synthesized in our laboratory.

[0129] AD7942: Fatty aldehyde decarbonylase, derived from Synechococcuselongatus PCC7942, the protein sequence is ac...

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Abstract

The invention relates to the field of biotechnology, especially to a gene and its application, a gene element, a method for synthesizing odd-numbered medium chain fatty aldehyde and a method for synthesizing even-numbered medium chain fatty hydrocarbon. The process of converting a precursor, namely fatty acid molecules, into an important intermediate metabolite--fatty aldehyde by the use of alpha-dioxygenase is an oxidation reaction. Extra reducing power and energy are provided without cells so as to reduce the burden of cell production. The technical limitation that fatty alcohol product chain length is only even number and fatty hydrocarbon product chain length is mostly odd number is broken. Bio-based bulk chemicals and bio-fuel molecules better match petroleum-based relative products.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to genes and their uses, gene elements, methods for synthesizing odd-numbered medium-chain aliphatic aldehydes, and methods for synthesizing even-numbered medium-chain aliphatic hydrocarbons. Background technique [0002] Medium-chain fatty alcohols have very important applications in industry due to their amphiphilic properties. They can be used in surfactants, medicine, cosmetics and energy fields, with a market value of 100-120 million US dollars. The aliphatic hydrocarbon molecules with a carbon chain length of 6-16 are the main components of aviation kerosene, which have the advantages of high calorific value, low vapor pressure, low freezing point and low hygroscopicity. 50% of commercial fatty alcohols are extracted from plant seeds or animal fats, and the remaining fatty alcohols and all hydrocarbons are refined from petroleum. No matter which method is used, it cannot meet the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N15/70C12N1/21C12P7/24C12P7/04C12P5/02
CPCC12N9/0006C12N9/0083C12N9/16C12N9/88C12P7/04C12P7/24C12Y101/01021C12Y401/99005C12P5/02
Inventor 元英进曹英秀刘夺丁明珠谢泽雄张金来
Owner TIANJIN UNIV
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